P-8

Oocyte cryopreservation success: A case study of an opportunity tosalvage an impending S IVF failure. D.E. Battaglia, P.E. Patton, D. Lee,M.J. Gorrill, K.A. Burry. Oregon Health and Science University, Portland,Oregon.

Background: Oocyte cryopreservation is an emerging technology that hasresulted in over 70 live births worldwide, mostly in the last two years. Withadvancements in freezing protocols, oocyte cryopreservation may presentan increasingly desirable option for a wide variety of patients.

Objective: To cryopreserve the oocytes of an IVF patient whose husbandsuffered from an acute medical emergency which prevented the collectionof a sperm sample for insemination during her cycle.

Design: A proven controlled-rate oocyte cryopreservation protocol wasutilized to cryopreserve the oocytes from an IVF cycle because sperm wereunavailable for insemination.

Materials and Methods: A 32 y.o. patient underwent controlled ovarianhyperstimulation for IVF. Twenty oocytes were obtained, but collection ofa sperm sample from her husband was medically contraindicated secondaryto a ruptured appendix. After her recovery from the oocyte retrieval thepatient agreed to cryopreserve her oocytes in an attempt to salvage hercycle. All 20 oocytes were frozen using the freezing protocol of Fabbri, etal. (Human Reprod. 16: 411–416, 2001). Three months later, all oocyteswere thawed and ICSI was performed.

Results: After thawing, 10/20 (50%) of the oocytes survived. Nine ofthese were at MII stage and were subjected to ICSI where 8/9 (89%)fertilized. Assessment of embryo development on d3 revealed embryos ofgood-fair quality. Three embryos were transferred on d3 and the remainingembryos were placed into extended culture. On d7 two embryos developedto expanded blastocysts of good quality. All remaining embryos werecryopreserved. Eleven days after embryo transfer the patient had a positivebhCG. Subsequent ultrasound examination revealed a twin clinical pregnancy.The twin status spontaneously reduced to an ongoing singleton pregnancy.

Conclusion: Oocyte cryopreservation was successful in salvaging whatcould have been an emotionally wrenching IVF failure due to an acutemedical situation which prevented sperm collection. Post-thaw oocyte sur-vival was lower than expected, however the surviving oocytes were of goodquality and ICSI fertilization was excellent. This case represents the firstfrozen oocyte pregnancy in the Northwest and provides encouraging datathat oocyte cryopreservation is a viable option for IVF patients underspecialized circumstances.

P-9

Body mass index of the egg recipient is predictive of success in an eggdonation program. V.L. Houserman, K.L. Honea, C.A. Long, K.E. Dal-ton, C.N. Gilbert, D.C. Merryman. ART Program of Alabama, Birming-ham, AL.

Background: A body mass index (BMI) of greater than 30 kg/m2 isconsidered obese while 25 to 30 is considered overweight. Many risks areassociated with obesity including menstrual disorders, infertility, surgicalcomplications, decreased success with fertility treatments and pregnancycomplications. BMI has been shown to have a negative effect on In VitroFertilization outcome using autologous eggs. The success of an egg dona-tion program is typically focused more on the characteristics of the eggdonor (ED) rather than the egg recipient (ER). Prior to cycling, the ER isgenerally scrutinized for the following: adequate endometrium, reasonablehealth status and acceptable screening test results relative to age. Strictcriteria for ER BMI are not usually emphasized.

Objective: To retrospectively compare the effect of ER BMI on theongoing/delivered pregnancy (ODP) rate in an egg donation program.

Materials and Methods: The data consisted of 127 ER cycles in which anembryo transfer (ET) was performed during the period January 1997through October 2002. Cycles utilizing testicular sperm (N � 7) wereexcluded from the data. Stimulation and embryo culture were carried out byconventional methods. Fisher’s exact test was used for statistical analysis.Statistical significance was defined as P � 0.05.

Results: Patients were grouped according to BMI: �25 and �25. TheODP rate per ET was 44.3% (35/79) when BMI was �25 and 25.0% (12/48)when BMI was �25 (P � 0.044, Table 1).

Table 1. Treatment outcome as a function of ER BMI in a group of 127cycles with ET.

BMIP�25 �25

Embryo transfers (ET) 79 48 -Ongoing � delivered

pregnancies (ODP)35 12 -

Ongoing � deliveredrate (OPD/ET)

44.3% 25.0% 0.044

Of note, BMI � 25, 25–30 and �30 yielded an ODP rate of 44.3% (35/79),28.6% (10/35) and 15.4% (2/13), respectively.

Conclusion:1) ER BMI � 25 resulted in a significantly higher delivery rate as

compared to ER BMI � 25.2) ER delivery rates appear to decrease as BMI increases.3) Weight loss to BMI � 25 should be strongly encouraged prior to

infertility treatment.

References:1) Loveland et al 2001 J Assist Reprod Genet Jul 18(7):382–6.2) Wittemer et al 2000 J Assist Reprod Genet Nov 17(10):547–52.3) Elkind-Hirsch et al 2001 Fertil Steril Apr 75(4):700–4.4) Salha et al 2001 Hum Fertil 4(1):37–42.

P-10

Evaluation of Mixed Protocols with Bravelle (hFSH) and Repronex(hMG) to Assess Clinical Efficacy (EMBRACE I) in Patients (18–33Years) Undergoing In Vitro Fertilization R.P. Marrs,1 J.H. Check,2 V.L.Schnell,3 M.H. Jacobs,4 M. Yemini,5 S.P. Oskowitz,6 J.H. Batzofin7 V.L.Houserman8, D.C. Marshall9 for the EMBRACE Study Group. 1CaliforniaFertility Partners, Santa Monica, CA; 2Cooper Institute for Reproductiveand Hormonal Disorders, Marlton, NJ; 3Center of Reproductive Medicine,Webster, TX; 4Fertility and IVF Center of Miami, Miami, FL; 5DiamondInstitute, Millburn, NJ; 6Boston IVF—The Surgery Center of Waltham,Waltham, MA; 7Huntington Reproductive Center, Pasadena, CA; 8ARTProgram, Birmingham, AL; 9Ferring Pharmaceuticals Inc., Suffern, NY

Principal Author: Dennis C. Marshall, Ph.D., Ferring Pharmaceuticals,Inc., 120 White Plains Road, Suite 400, Tarrytown, NY 10591. Phone:914.333.8985. Fax: 914.631.5120. Email: marshall@ferringusa.com

Introduction: There is widespread use of combined FSH and hMG (mixedprotocols) in controlled ovarian hyperstimulation (COH) for in vitro fertil-ization (IVF). However, there are no systematic efficacy and safety studiesexamining various ratios of FSH and hMG mixed in a single daily dose.

Objective: To assess the therapeutic efficacy and safety of continuous andsequential dose ratios of FSH:hMG (Bravelle, highly purified FSH andRepronex, hMG) combined in the same syringe and administered subcuta-neously, once daily, in COH-IVF patients 18–33 years of age.

Materials and Methods: Eligible patients received leuprolide acetate (LA,0.5 mg, OD, SC) starting seven days before onset of menses and continuedfor ≤ 20 days until estradiol (E2) was ≤40 pg/ml withendometrial lining ≤6mm. Thereafter, LA was reduced to 0.25 mg/dand continued until the day before hCG administration. Patients wererandomized to treatment groups A, B or C, and gonadotropin stimulationbegan for ≤15 days. The FSH:hMG vial ratios were: Group A had a1:1 ratio throughout; Group B began with FSH only then changed to andmaintained a 1:1 ratio after day 5 of gonadotropins; Group C had a 2:1 ratiothat, after day 5 of gonadotropins, was sequentially adjusted to 3:1, 4:1 or5:1 as needed to maximum FSH dose of 450 IU. When ultrasound showed≥3 follicles with diameters of ≥16 mm, and acceptable E2levels, gonadotropins were stopped and hCG (10,000 IU IM) was admin-istered; oocytes were retrieved 34–36 hrs later. Primary efficacy was thenumber of oocytes retrieved.

Results: Oocyte yield and pregnancy rates were excellent for all treatmentgroups. Mean daily peak serum E2 levels were significantly greater andFSH doses were significantly less in Group A vs Group B or Group C. Therewere no differences in adverse events.

S14 PCRS Abstracts Vol. 79, Suppl. 2, April 2003

Patient Demographics, Efficacy Results and Safety Data: EMBRACE I

ParameterTreatment A

(n � 35)Treatment B

(n � 39)Treatment C

(n � 34) P-value

Age(mean � SD;yrs)

30.0 (2.2) 30.0 (2.3) 30.5 (2.0) 0.527

BMI (kg/m2) 24.2 (3.7) 24.2 (4.4) 24.3 (4.7) 0.997Duration of

treatment(mean days� SD)

9.4 (1.7) 9.4 (1.2) 9.9 (1.4) 0.272

Total gonadotropindose (mean IU� SD)

2167.5 (690.9) 2319.2 (551.2) 2616.2 (687.3) 0.015

Peak serum E2levels(mean � SD;pg/ml)

2988.4 (1599.9) 1908.2 (1058.2) 2222.0 (780.7) 0.001

Patients with oocyteretrieval andembryotransfer (%)

32 (94.1) 34 (87.2) 31 (93.9) 0.509

Total oocytesretrieved(mean � SD)

16.7 (6.2) 13.6 (9.3) 15.5 (10.2) 0.306

Mature oocytes(mean � SD)

12.3 (6.2) 10.7 (7.7) 11.6 (10.0) 0.683

Oocytes withnormalfertilization(mean � SD)

9.7 (5.6) 7.7 (7.0) 9.1 (7.7) 0.423

Embryos transferred(mean � SD)

2.8 (0.9) 2.5 (1.2) 2.7 (1.0) 0.385

Implantation rate(%)

28.1 32.0 28.3 0.804

Patients withchemicalpregnancy (%)

18 (52.9) 17 (43.6) 20 (58.8) 0.502

Patients withclinicalpregnancy (%)

16 (47.1) 17 (43.6) 16 (47.1) 0.948

Patients withcontinuingpregnancy (%)

16 (47.1) 16 (41.0) 15 (44.1) 0.852

Patients with anyadverseevent (%)

19 (54.3) 19 (48.7) 25 (73.5) 0.084

Patients with seriousadverse events(%)

2 (5.7) 2 (5.1) 1 (2.9) 1.000

Conclusion: The same-syringe combination and administration of thesehuman-derived FSH and hMG hormones resulted in excellent oocyte yield,implantation and pregnancy rates. In patients 18–33 years, FSH:hMGstarting ratios of 1:1 produced statistically significantly greater E2 responsesand required significantly less total dose FSH than the other ratios tested.

P-11

Early pregnancy hCG differences associated with the timing of embryotransfer following in vitro fertilization. B.S. Shapiro,1,2 K.S. Richter,1

S.T. Daneshmand.1,2 1Fertility Center of Las Vegas and 2Dept of OB/GYN,University of Nevada School of Medicine, Las Vegas, NV.

Background: Early pregnancy serum hCG is indicative of embryo im-plantation and pregnancy outcome.

Objective: The association between the timing of embryo transfer fol-lowing in vitro fertilization (IVF) and early pregnancy hCG concentrationswas examined.

Materials and Methods: Serum hCG concentrations (standardized to day15 after oocyte retrieval and adjusted for patient weight) of all pregnant IVFpatients at a private ART center from January 1996 through February 2002were retrospectively investigated. Comparisons were made between cleav-age stage and blastocyst stage embryo transfers, and between day 5 and day6 blastocyst transfers. Embryos transferred prior to October 1997 weretransferred on day 3 after oocyte retrieval (cleavage stage). Thereafter,transfers were conducted on either day 5 or day 6 after oocyte retrievalaccording to the day of blastocyst expansion. Implantations were defined by

ultrasound identification of gestational sacs and fetal heart motion. SerumhCG titers per subsequently identified implantation (natural log trans-formed) were compared between patient groups by t-test. ANCOVA, in-cluding the (transformed) number of implantations as a covariate, was alsoused to examine the association between the time of embryo transfer and(transformed) hCG titers on day 15 after retrieval.

Results: Statistical comparisons indicated significant differences (p �0.005) in serum hCG according to the day of blastocyst expansion andtransfer, suggesting a delay of approximately 12.5 to 13 hours in hCGproduction for the later expanding blastocysts. Comparisons between cleav-age stage and blastocyst transfers indicated significant differences (p �0.05) in day 15 hCG titers, suggesting a delay of approximately 8 to 8.5hours in hCG production for pregnancies resulting from blastocyst ascompared to cleavage stage embryo transfers.

Conclusions: A delay in blastocyst expansion is associated with a reduc-tion in serum hCG concentration (implying delayed implantation) if preg-nancy occurs. Comparisons of serum hCG concentrations between pregnan-cies resulting from cleavage or blastocyst stage embryo transfers suggestthat prolonged exposure to in vitro conditions may result in a small butsignificant delay in embryo implantation, or a reduction in trophoblastic tissue.

P-12

Granulocyte-macrophage colony-stimulating factor enhances humanembryo development to the blastocyst stage: a randomized study. B.S.Shapiro,1,2 K.S. Richter,1 S.T. Daneshmand,1,2 P. Quinn,3 B. Behr.4 1Fer-tility Center of Las Vegas, 2Dept of OB/GYN, University of Nevada Schoolof Medicine, Las Vegas, NV, 3SAGE BioPharma, San Clemente, CA, 4Deptof OB/GYN, Stanford University School of Medicine, Stanford, CA.

Background: Currently available sequential media lack growth factors,although pre-implantation embryos are naturally exposed to many growthfactors in vivo.

Objective: To assess the value of supplementing sequential embryo culturemedia with granulocyte-macrophage colony-stimulating factor (GM-CSF) onembryo development from the 2pn stage to the blastocyst stage.

Design: Prospective, randomized, blinded study comparing media that areidentical except for the presence or absence of granulocyte-macrophagecolony-stimulating factor.

Materials and Methods: Patients having two or more 2pn oocytes follow-ing IVF were enrolled following informed consent (n � 72). For eachpatient, all 2pn oocytes were randomly divided between GM-CSF andcontrol treatments. Embryos were cultured to the blastocyst stage in Quinn’sAdvantage Protein Plus sequential media (SAGE BioPharma) with or with-out GM-CSF supplementation. One to three blastocysts were transferred topatients on day 5 or 6 after retrieval. Cell numbers at 48 and 72 hours andnumbers of expanded blastocysts were compared by paired t-test. Propor-tions of 2pn embryos to cleavage stage, cleaving embryos to compactionstage, compacting embryos to cavitation stage, and cavitating embryos tothe expanded blastocyst stage were compared by Wilcoxon signed rank test.This protocol was approved by our local IRB before initiation of the study.

Results: Supplementation of media with GM-CSF resulted in greater cellnumbers per embryo at 72 hours (6.1 vs 5.8, p � 0.047), and more expandedblastocysts per 2pn oocyte (p � 0.0005), and per cleaving embryo (p �0.002) or compacting embryo (p � 0.028). GM-CSF supplementationresulted in a 50% increase in the number of expanded blastocysts availablefor transfer (1.6 vs 1.1, p � 0.001). Sixty-six patients (92%) underwentembryo transfer. Pregnancy and implantation rates (Table) were similarregardless of the presence or absence of GM-CSF in the culture media.

Pregnancy and implantation rates according to the culture media oftransferred embryos

TransferredEmbryos

Pregnancy/Transfer

Implantation/Embryo

1–3 embryos, all GM-CSF 8/17 (47%) 10/31 (32%)1–3 embryos, all Non-GM-CSF 2/4 (50%) 2/8 (25%)3 embryos, 2 GM-CSF & 1 Non-GM-CSF 7/18 (39%) 11/54 (20%)3 embryos, 1 GM-CSF & 2 Non-GM-CSF 1/2 (50%) 1/6 (17%)2 embryos, 1 GM-CSF & 1 Non-GM-CSF 11/25 (44%) 14/50 (28%)

Conclusions: Supplementation of sequential media with the growth factorGM-CSF enhances human embryo growth throughout pre-implantation

FERTILITY & STERILITY� S15